Compositions and methods of treating abnormal cell proliferation

ABSTRACT

A composition is described comprising a vitamin D analog and a retinoid wherein: (a) the vitamin D analog is capable of binding a vitamin D receptor or being converted in vivo into a compound capable of binding a vitamin D receptor; and (b) the retinoid is selected from the group consisting of retinol in a concentration of at least about 1.0% and a retinoid characterized by having a substitution at the 4-position. Further, methods of treating disorders characterized by abnormal cell-proliferation and/or cell-differentiation are also described.

RELATED APPLICATIONS

This application is a continuation-in-part of U.S. Ser. No. 09/351,020,filed Jul. 12, 1999 now U.S. Pat. No. 6,242,435, which is acontinuation-in-part of U.S. Ser. No. 09/116,632, filed Jul. 16, 1998abandoned.

Throughout this application, various references are referred to withinparentheses. Disclosures of these publications in their entireties arehereby incorporated by reference into this application to more fullydescribe the state of the art to which this invention pertains.

FIELD OF THE INVENTION

The present invention relates to compositions comprising certainretinoids and vitamin D analogs useful in inducing differentiation andinhibiting undesirable proliferation of cells, such as cancer cells andskin cells. The present invention also relates to methods of using theabove compositions in the treatment of diseases and conditionscharacterized by abnormal cell differentiation and/or cellproliferation.

DESCRIPTION OF THE RELATED ART

Abnormal cell differentiation and/or cell differentiation is associatedwith many conditions and diseases. For instance, hyperproliferation ofepithelial cells is associated with psoriasis which causes the skin toshed itself too rapidly, every three to four days. The goal in treatingpsoriasis is to reduce inflammation and to slow down rapid skin celldivision.

U.S. Pat. No. 4,866,048 discloses that certain vitamin D derivatives, inparticular calcitriol (1 alpha,25-dihydroxy-vitamin D₃ or) andcalcipotriol are able to stimulate the differentiation of cells andinhibit excessive cell proliferation, and it has been suggested thatthese compounds are useful in the treatment of diseases characterized byabnormal cell differentiation and/or cell differentiation such asleukemia, myelofibrosis, psoriasis and acne.

Certain retinoids are also known for their antiproliferative anddifferentiation activity. For instance, retinol (vitamin A) is anendogenous compound which occurs naturally in the human body and isessential for normal cell differentiation of certain cell types such asepithelial cells. Retinoic acid is believed to be an active derivativeof retinol. Thus, retinoic acid is believed to be more effective thanretinol and retinyl esters at providing skin benefits.

Natural and synthetic vitamin A derivatives (including retinoic acid)have been used extensively in the treatment of a variety of skin andhyperproliferation disorders. For example, retinoic acid has beenemployed to treat certain types of leukemia like acute apromyelocyticleukemia as well as a variety of skin conditions such as acne, wrinkles,psoriasis, age spots and discoloration (Vahlquist, A. et al., J. Invest.Dermatol., Vol. 94, Holland D. B. and Cunliffe, W. J. (1990), pp.496-498; Ellis C. N. et al., “Pharmacology of Retinols in Skin”, Vasel,Karger, Vol. 3, (1989), pp.249-252; Lowe, N. J. et al., Vol. 3, (1989),pp. 240-248; PCT Patent Application No. WO 93/19743). Although retinoidshave been viewed classically as cancer prevention agents, considerablelaboratory evidence supports their testing as antitumor drugs as well(Cancer Treat Rep 1987; 71: 493-515 May, 1987).

It is important to note that while clinical experience with eitherretinoids or vitamin D derivatives against conditions associated withabnormal cell differentiation and/or cell differentiation has met withcertain amount of success in some instances, these compounds havefrequently been unable to provide the desired clinical results.

For instance, the synthetic Vitamin D, calcipotriol, or retinoic acidwhich are available in prescription form are somewhat useful forindividuals with localized psoriasis. However, these compound are notvery effective on most patients.

Therapeutic regimens for acne involve local and systemic therapies,although the former is indicated in the vast majority of cases. Topicalapplication of a variety of chemical application which include mainlysulfur, resorcinol, salicylic acid, benzoyl peroxide, and retinoic acidare frequently used to treat acne. All the foregoing agents are known as“peeling” or “drying” agents which are believed to exert theirtherapeutical effect by causing erythema, irritation, and desquaminationof the skin to expel comedones. The therapeutic efficacy of theseagents, however, is rather variable, and their utility is limitedpartially because of the irritation caused by their application (seeU.S. Pat. No. 3,932,665). Oral formulations of retinoic acid are alsoused but serious side effects are associated with the oral use of thiscompound including severe fetal malformation in pregnant women.

SUMMARY OF THE INVENTION

The present invention provides compositions comprising certain vitamin Dand retinoid compounds which are useful for the treatment of disorderscharacterized by abnormal cell-proliferation and/orcell-differentiation.

Specifically, the present invention provides a composition comprising avitamin D analog and a retinoid, wherein:

(a) the vitamin D analog is capable of binding a vitamin D receptor orbeing converted in vivo into a compound capable of binding a vitamin Dreceptor; and

(b) the retinoid is selected from the group consisting of retinol in aconcentration of at least about 1.0% by weight, a compound in aconcentration of at least about 1.0% by weight capable of beingconverted in vivo into retinol, retinoid D with an alcohol CH₂OHterminal side chain, retinoid D with an ester at the terminal sidechain, retinoid D with an ether at the terminal side chain, retinoid Dwith an aldehyde at the terminal side chain, and retinoid D with acarboxylic acid at the terminal side chain, wherein retinoid D with thealcohol CH₂OH terminal side chain has the structure:

wherein the configuration at the 7-, 9-, 11- and 13-position doublebonds is independently Z or E and wherein R₁ is selected from the groupconsisting of

wherein the keto group at the 4-position is free or protected, or isreplaced by a thioketone group which is free or protected or is replacedby C₁₋₆-alkylidene group;

wherein X is selected from the group consisting of hydrogen andC₁₋₆-alkyl and Y is selected from the group consisting of C₁₋₆ -alkyl,hydroxyl, alkoxyl, acyloxyl, halide, azide, sulfhydryl, amine andC₁₋₆-alkyl substituted amino and wherein the absolute configuration atthe 4-position is independently R or S;

wherein X₁, Y₁ are independently selected from the group consisting ofhydrogen, C₁₋₆-alkyl, hydroxyl, alkoxyl, acyloxyl, halide, azide,sulfhydryl, amine and C₁₋₆-alkyl substituted amino and Z₁ is selectedfrom the group consisting of C₁₋₆-alkyl, hydroxyl, alkoxyl, acyloxyl,halide, azide, sulfhydryl, amine and C₁₋₆-alkyl substituted amino;

wherein X₂ is selected from the group consisting of hydrogen,C₁₋₆-alkyl, hydroxyl, alkoxyl, acyloxyl, halide, azide, sulfhydryl,amine and C₁₋₆-alkyl substituted amino and Z₂ is selected from the groupconsisting of C₁₋₆-alkyl, hydroxyl, alkoxyl, acyloxyl, halide, azide,sulfhydryl, amine and C₁₋₆-alkyl substituted amino;

wherein X₃ and Y₃ are independently selected from the group consistingof hydrogens, C₁₋₆-alkyl, hydroxyl, alkoxyl, acyloxyl, halide, azide,sulfhydryl, amine and C₁₋₆-alkyl substituted amino so long as X₃ and Y₃are not both hydrogens.

The present inventions also provides methods for treating variousconditions associated with abnormal cell proliferation and/or abnormalcell differentiation.

DETAILED DESCRIPTION OF THE INVENTION

In accordance with the invention, It has been surprisingly discoveredthat a composition comprising a vitamin D analog and a certain retinoidis useful in treating a subject suffering from a disorder characterizedby abnormal cell-proliferation and/or cell-differentiation moreeffectively than either a composition comprising a vitamin D or theabove retinoid or a composition comprising a vitamin D analog with othertypes of retinoids.

The retinoid used in the composition of the present invention isselected from the group consisting of retinol in a concentration of atleast about 1.0% by weight, a compound in a concentration of at leastabout 1.0% by weight capable of being converted in vivo into retinol,retinoid D with an alcohol CH₂OH terminal side chain, retinoid D with anester at the terminal side chain, retinoid D with an ether at theterminal side chain, retinoid D with an aldehyde at the terminal sidechain, and retinoid D with a carboxylic acid at the terminal side chain,wherein retinoid D with the alcohol CH₂OH terminal side chain has thestructure:

wherein the configuration at the 7-, 9-, 11- and 13-position doublebonds is independently Z or E and wherein R₁ is selected from the groupconsisting of

wherein the keto group at the 4-position is free or protected, or isreplaced by a thioketone group which is free or protected or is replacedby C₁₋₆-alkylidene group;

wherein X is selected from the group consisting of hydrogen andC₁₋₆-alkyl and Y is selected from the group consisting of C₁₋₆-alkyl,hydroxyl, alkoxyl, acyloxyl, halide, azide, sulfhydryl, amine andC₁₋₆-alkyl substituted amino and wherein the absolute configuration atthe 4-position is independently R or S;

wherein X₁, Y₁ are independently selected from the group consisting ofhydrogen, C₁₋₆-alkyl, hydroxyl, alkoxyl, acyloxyl, halide, azide,sulfhydryl, amine and C₁₋₆-alkyl substituted amino and Z₁ is selectedfrom the group consisting of C₁₋₆-alkyl, hydroxyl, alkoxyl, acyloxyl,halide, azide, sulfhydryl, amine and C₁₋₆-alkyl substituted amino;

wherein X₂ is selected from the group consisting of hydrogen,C₁₋₆-alkyl, hydroxyl, alkoxyl, acyloxyl, halide, azide, sulfhydryl,amine and C₁₋₆-alkyl substituted amino and Z₂ is selected from the groupconsisting of C₁₋₆-alkyl, hydroxyl, alkoxyl, acyloxyl, halide, azide,sulfhydryl, amine and C₁₋₆-alkyl substituted amino;

wherein X₃ and Y₃ are independently selected from the group consistingof hydrogens, C₁₋₆-alkyl, hydroxyl, alkoxyl, acyloxyl, halide, azide,sulfhydryl, amine and C₁₋₆-alkyl substituted amino so long as X₃ and Y₃are not both hydrogens.

Preferably, the abnormal cell proliferation treated with the compositionof the present invention is associated with cancer cells and morepreferably with skin cancer such as melanoma. Also more preferably, theabnormal cell proliferation is associated with cancer cells that can atleast partially respond to hormone or retinoid treatment.

The present invention also provides a method of treating a subjectsuffering from a disorder selected from the group consisting ofpsoriasis, acne, atopic dermatitis, eczema, rosacea, actinic keratosis,seborrheic dermatitis, and congenital keratinization disorders, in whichany composition of the present invention is administered to the subjectin need of such treatment. Preferably, the disorder is psoriasis,eczema, or acne.

The present invention further provides a method of treating one or moreconditions of the skin selected from the group consisting of dry skin,photodamaged skin, age spots, aged skin, increasing stratum corneumflexibility, wrinkles, fine lines, actinic blemishes, skin dyschromias,and ichthyosis, comprising applying to the skin having said one or morecondition any composition of the present invention. Preferably, the skincondition is actinic blemishes or fine wrinkles.

The present invention also provides methods for preventing or treatingindividuals suffering hair loss such as male pattern baldness or femalepattern baldness comprising applying to the affected areas of the skinany composition of the present invention.

The present invention further provides methods for restoring the naturalcolor of gray hair comprising applying to the affected areas of the skinany composition of the present invention.

For the purpose of this invention, treatment of a subject with thecomposition of the present invention means administering or applying tosaid subject either together or separately both the vitamin D analog andthe retinoid as defined in the composition of the present invention.Thus, the vitamin D analog and the retinoid may be administered orapplied together or separately using either the same or different formsof administration or application.

Also, for the purpose of this invention, the term “vitamin D analog” isdefined as a compound capable of binding a vitamin D receptor (notnecessarily all) or being converted in vivo into a compound capable ofbinding a vitamin D receptor (not necessarily all). The term “vitamin Danalog” includes but is not limited to vitamin D₂ and vitamin D₃derivatives such as cholecalciferol, calcifediol, calcitriol,calcipotriol, ergosterol, ergocalciferol, dihydrotachysterol,1,25-dihydroxyergocalciferol, 25-hydroxydihydrotachysterol, and thevitamin D analogs disclosed in U.S. Pat. No. 4,866,048. Preferredanalogs are cholecalciferol, calcifediol, calcitriol, calcipotriol andthe vitamin D analogs disclosed in U.S. Pat. No. 4,866,048. Morepreferred analogs are cholecalciferol, calcifediol, calcitriol andcalcipotriol. Most preferred analogs are calcitriol and calcipotriol.

The concentration of the vitamin D analog may vary from about 0.0001% toabout 10% by weight of the total composition of the invention.Preferably, the concentrations employed of vitamin D analogs that candirectly bind to the vitamin D receptors, range from about 0.0001% toabout 1%, more preferably from about 0.0005% to about 0.05%, still morepreferably from about 0.009% to about 0.5%, yet still more preferablyfrom about 0.001 to about 0.008%, and most preferably at about 0.005%.

Preferably, the concentration employed of vitamin D analogs that can beconverted in vivo to a compound capable of binding a vitamin D receptoris from about 0.001% to about 10%, more preferably from about 0.01% toabout 8%, still more preferably from about 1% to about 6%, and mostpreferably from about 2% to about 5%.

Retinoid D is preferably 4-oxo-retinoic acid, 4-oxo-retinol, and4-oxo-retinal, 4-hydroxy-retinol, 4-hydroxy-retinal, 4-oxo-retinylester, and 4-hydroxyretinyl ester. The most preferred retinoid is4-oxo-retinol. Preferably, the concentration of retinoid D in thecompositions of the invention ranges from about 0.001% to about 1%, morepreferably from about 0.025% to about 0.1%, most preferably about 0.05%.

Also, for the purpose of this invention, the term “retinol” includes butis not limited to the following: all-trans-retinol, 13-cis-retinol,1-cis-retinol, 9-cis-retinol, 3,4-didehydro-retinol. Preferred isomersare all-trans-retinol, 13-cis-retinol, 3,4-didehydro-retinol, and9-cis-retinol. Most preferred is all-trans-retinol due to its widecommercial availability. The concentration employed of retinol is atleast about 0.1%, preferably at least 0.3% by weight of the total weightof the composition. More preferably, the concentration such retinoid isfrom about at 0.3% to about 20%, more preferably from about 0.5% toabout 15%, still more preferably from about 0.5% to about 10%, stillmore preferably from about 1% to about 10%, still more preferably fromabout 2% to about 10%, and most preferably about 5%.

The retinoids that can be converted in vivo to retinol include but arenot limited to retinyl esters, retinyl-glucoronides, retinal,3,4-didehydro-retinol. Compounds that are converted spontaneously byisomerization are also included in the compounds of the invention.

Retinyl ester is an ester of retinol and is capable of being convertedin vivo into retinol. Retinyl esters suitable for use in the presentinvention include but are not limited to C₁-C₃₀ esters of retinol,preferably C₂-C₂₀ esters, and most preferably C₂, C₃, and C₆ estersbecause they are commonly available. Examples of retinyl, esters includebut are not limited to: retinyl palmitate, retinyl formate, retinylacetate, retinyl propionate, retinyl butyrate, retinyl valerate, retinylisovalerate, retinyl hexanoate, retinyl heptanoate, retinyl octanoate,retinyl nonanoate, retinyl decanoate, retinyl undecandate, retinyllaurate, retinyl tridecanoate, retinyl myristate, retinylpentadecanoate, retinyl heptadecanoate, retinyl stearate, retinylisostearate, retinyl nonadecanoate, retinyl arachidonate, retinylbehenate, retinyl linoleate, and retinyl oleate.

The preferred retinyl esters for use in the present invention areretinyl palmitate, retinyl acetate, retinyl propionate and retinyllinoleate. More preferred retinyl esters are retinyl palmitate andretinyl acetate, the most preferred retinyl ester is retinyl palmitate.

The concentration employed of the retinoid that can be converted in vivoto retinol is at least about 0.1%, preferably at least 0.3% by weight ofthe total weight of the composition. More preferably, the concentrationsuch retinoid is from about at 0.3% to about 20%, more preferably fromabout 0.5% to about 15%, still more preferably from about 0.5% to about10%, still more preferably from about 1% to about 10%, still morepreferably from about 2% to about 10%, and most preferably about 5%.

It has also been surprisingly discovered that a composition comprisingretinol in a concentration of at least about 1.5% or a compound in aconcentration of at least about 1.5% capable of being converted in vivointo retinol is as effective as retinoic acid in treating one or moreconditions of the skin selected from the group consisting of dry skin,photodamaged skin, age spots, aged skin, increasing stratum corneumflexibility, wrinkles, fine lines, actinic blemishes, skin dyschromias,ichthyosis and acne. The term “retinol” and the compounds capable ofbeing converted into retinol has been defined above. Preferably, forthis composition, the concentration employed of retinol or of thecompound capable of being converted in vivo into retinol is at leastabout 1.8%, more preferably at least about 2% by weight of the totalweight of the composition. More preferably, the concentration suchretinoid is from about at 2% to about 20%, more preferably from about 2%to about 15%, still more preferably from about 2% to about 10%, and mostpreferably about 5%.

The compositions of the present invention are preferably topical and/orpharmaceutical. They may be in the form of a cream, ointment, and gel.They may also comprise a cosmetically acceptable vehicle to act as adiluent, dispersant or carrier for the active components in thecomposition, so as to facilitate their distribution when the compositionis applied to the skin.

Vehicles other than water can include liquid or solid emollients,solvents, humectants, thickeners and powders. An especially preferrednonaqueous carrier is a polydimethyl siloxane and/or a polydimethylphenyl siloxane. Silicones of this invention may be those withviscosities ranging anywhere from about 10 to 10,000,000 centistokes at25° C. Especially desirable are mixtures of low and high viscositysilicones. These silicones are available from the General ElectricCompany under trademarks Vicasil, SE and SF and from the Dow CorningCompany under the 200 and 550 Series. Amounts of silicone which can beutilized in the compositions of this invention range anywhere from 5% to95%, preferably from 25% to 90% by weight of the composition.

The cosmetically acceptable vehicle will usually form from 5% to 99.9%,preferably from 25% to 80% by weight of the emulsion, and can, in theabsence of other cosmetic adjuncts, form the balance of the composition.

An oil or oily material may be present in the claimed compositions,together with an emulsifier to provide either a water-in-oil emulsion oran oil-in-water emulsion, depending largely on the averagehydrophilic-lipophilic balance (HLB) of the emulsifier employed.

Various types of active ingredients may be present in cosmeticcompositions of the present invention. Various types of activeingredients may be present in cosmetic compositions of the presentinvention. Actives are defined as skin benefit agents other thanemollients and other than ingredients that merely improve the physicalcharacteristics of the composition. Although not limited to thiscategory, general examples include sunscreens and tanning agents.

Sunscreens include those materials commonly employed to blockultraviolet light. Illustrative compounds are the derivatives of PABA,cinnamate and salicylate. For example, octyl methoxycinnamate and2-hydroxy-4-methoxy benzophenone (also known as oxybenzone) can be used.Octyl methoxycinnamate and 2-hydroxy-4-methoxy benzophenone arecommercially available under the trademarks, Parsol MCX andBenzophenone-3, respectively. The exact amount of sunscreen employed inthe emulsions can vary depending upon the degree of protection desiredfrom the sun's UV radiation.

Another preferred optional ingredient is selected from essential fattyacids (EFAs), i.e., those fatty acids which are essential for the plasmamembrane formation of all cells (in keratinocytes, EFA deficiency makescells hyperproliferative). Supplementation of EFA corrects this. EFAsalso enhance lipid biosynthesis of epidermis and provide lipids for thebarrier formation of the epidermis. The essential fatty acids arepreferably chosen from linoleic acid, gamma-linolenic acid,homo-gamma-linolenic acid, columbinic acid, eicosa-(n-6,9,13)-trienoicacid, arachidonic acid, gamma-linolenic acid, timnodonic acid, hexaenoicacid and mixtures thereof.

Emollients are often incorporated into cosmetic compositions of thepresent invention. Levels of such emollients may range from about 0.5%to about 50%, preferably between about 5% and 30% by weight of the totalcomposition. Emollients may be classified under such general chemicalcategories as esters, fatty acids and alcohols, polyols andhydrocarbons.

Esters may be mono- or di-esters. Acceptable examples of fatty di-estersinclude dibutyl adipate, diethyl sebacate, diisopropyl dimerate, anddioctyl succinate. Acceptable branched chain fatty esters include2-ethyl-hexyl myristate, isopropyl stearate and isostearyl palmitate.Acceptable tribasic acid esters include triisopropyl trilinoleate andtrilauryl citrate. Acceptable straight chain fatty esters include laurylpalmitate, myristyl lactate, oleyl eurcate and stearyl oleate. Preferredesters include coco-caprylate/caprate (a blend of coco-caprylate andcoco-caprate) , propylene glycol myristyl ether acetate, diisopropyladipate and cetyl octanoate.

Suitable fatty alcohols and acids include those compounds having from 10to 20 carbon atoms. Especially preferred are compounds such as cetyl,myristyl, palmitic and stearyl alcohols and acids.

Among the polyols which may serve as emollients are linear and branchedchain alkyl polyhydroxyl compounds. For example, propylene glycol,sorbitol and glycerin are preferred. Also useful may be polymericpolyols such as polypropylene glycol and polyethylene glycol. Butyleneand propylene glycol are also especially preferred as penetrationenhancers.

Exemplary hydrocarbons which may serve as emollients are those havinghydrocarbon chains anywhere from 12 to 30 carbon atoms. Specificexamples include mineral oil, petroleum jelly, squalene andisoparaffins.

Another category of functional ingredients within the cosmeticcompositions of the present invention are thickeners. A thickener willusually be present in amounts anywhere from 0.1 to 20% by weight,preferably from about 0.5% to 10% by weight of the composition.Exemplary thickeners are cross-linked polyacrylate materials availableunder the trademark Carbopol from the B. F. Goodrich Company. Gums maybe employed such as xanthan, carrageenan, gelatin, karaya, pectin andlocust beans gum. Under certain circumstances the thickening functionmay be accomplished by a material also serving as a silicone oremollient. For instance, silicone gums in excess of 10 centistokes andesters such as glycerol stearate have dual functionality.

Powders may be incorporated into the cosmetic composition of theinvention. These powders include chalk, talc, Fullers earth, kaolin,starch, smectite clays, chemically modified magnesium aluminum silicate,organically modified montmorillonite clay, hydrated aluminum silicate,fumed silica, aluminum starch octenyl succinate and mixtures thereof.

Other adjunct minor components may also be incorporated into thecosmetic compositions. These ingredients may include coloring agents,opacifiers, perfumes and preservatives (e.g., imidazolidinyl urea,dimethyl imidazolidinone and diazolidinyl urea). Amounts of thesematerials may range anywhere from 0.001% up to 20% by weight of thecomposition.

The composition according to the invention is intended primarily but notexclusively as a product for topical application to human skin and as aproduct to modulate cell differentiation. In use, a small quantity ofthe composition, for example from 1 to 5 ml, is applied to exposed areasof the skin, from a suitable container or applicator and, if necessary,it is then spread over and/or rubbed into the skin using the hand orfingers or a suitable device.

The topical skin treatment composition of the invention can beformulated as a lotion having a viscosity of from 4,000 to 10,000 mPas,a fluid cream having a viscosity of from 10,000 to 20,000 mPas or acream or a gel having a viscosity of from 20,000 to 100,000 mPas orabove. The composition can be packaged in a suitable container to suitits viscosity and intended use by the consumer. For example, a lotion orfluid cream can be packaged in a bottle or a roll-ball applicator, or acapsule, or a propellant-driven aerosol device or a container fittedwith a pump suitable for finger operation. When the composition is acream, it can simply be stored in a non-deformable bottle or squeezecontainer, such as a tube or a lidded jar.

The invention accordingly also provides a closed container containing acosmetically acceptable composition as herein defined.

The following specific examples further illustrate the invention, butthe invention is not limited thereto.

EXAMPLE 1

This example compares therapeutically applied retinyl palmitate,retinoic acid, calcitriol, and combinations thereof in a cream atdifferent concentrations with the effectiveness of the cream without anyof the above compounds present in treating acne. The base cream used toprepare the experimental formulations is the commercially availableLUBRIDERM cream. The different compounds at different concentrationswere added to the cream and mixed very well. Sixty six volunteers wererecruited and were randomly assigned to each of the groups. the subjectswere selected on the basis of their having moderate to severepapular-pustular acne. Each group consisted of 3 males and 3 females. Noother acne treatment was permitted during the period. Preparations wereapplied to the face in the morning and evening after washing the facewith ordinary soap. Observations were made at time 0, 1, 4, and 8 weeksto assure that treatment was carried out according to direction.Judgments of “worse”, “no change”, “mild”, and “good” were made after 8weeks of treatment. Table 1 illustrates the results.

TABLE 1 Results of treatment with various compounds on acne no Treatmentworse change mild good Control 2 4 0 0 0.1% retinoic acid 0 2 3 1 0.1%retinyl 2 3 1 0 palmitate 1% retinyl palmitate 0 4 2 0 1.5% retinyl 0 13 2 palmitate 5% retinol 0 1 2 3 10% retinol 0 0 3 3 0.0025% calcitriol2 3 1 0.1% retinoic acid 0 1 3 2 and 0.0025% calcitriol 0.1% retinyl 1 22 1 palmitate and 0.0025% calcitriol 0.5% retinyl 0 1 3 2 palmitate and0.0025% calcitriol 5% retinyl palmitate 0 0 1 5 and 0.0025% calcitriol0.1% 4-oxo-retinol 0 1 4 1 0.1% 4-oxo-retinol 0 0 3 3 and 0.0025%calcitriol

Table 1 illustrates that retinyl palmitate in concentrations at about1.5% and more shows a remarkable improvement over lower concentrationsin treating acne. In addition, the combination of 4-oxo-retinol andcalcitriol shows a synergistic effect when compared with either4-oxo-retinol and calcitriol alone. A synergistic effect is also seenwhen calcitriol is combined with retinyl palmitate particularly atconcentrations of 0.5% of retinyl palmitate.

The use of each of retinoic acid and calcitriol caused skin irritationwhile the use of retinyl palmitate or 4-oxo-retinol did not.

EXAMPLE 2

This example compares therapeutically applied retinyl palmitate,retinoic acid, calcitriol, and combinations thereof in a cream atdifferent concentrations with the effectiveness of the cream without anyof the above compounds present in treating psoriasis. The base creamused to prepare the experimental formulations is the commerciallyavailable LUBRIDERM cream. The different compounds at differentconcentrations were added to the cream and thoroughly mixed. Also,commercially available 0.005% calcipotriol (DOVONEX) and commerciallyavailable 0.005% calcipotriol was supplemented with 5% retinyl palmitateand employed in treating psoriasis. Two different and distant psoriaticspots were selected on the skin of patients diagnosed with psoriasis fordifferent treatment. Each patient used two type of creams twice a day,one cream on each selected spot.

Group I consisting of five patients applied on one selected spot (spotA) control LUBRIDERM cream and on the other selected spot (spot B) creamcontaining 0.1% retinoic acid. None of the patients showed anyimprovement in either spots even after 12 weeks of treatment.

Group II consisting of five patients applied on one selected spot (spotA) control LUBRIDERM cream and on the other selected spot (spot B) creamcontaining 0.1% retinyl palmitate. No change in spot A was observedafter 4 weeks of treatment. Three out of five patients showed mildimprovement in itching after 3 days in spot B but no improvement instopping scaling which results from cellular hyperproliferation.

Group III consisting of five patients applied on one selected spot (spotA) control LUBRIDERM cream and on the other selected spot (spot B) creamcontaining 1% retinyl palmitate. No change in spot A was observed after4 weeks of treatment. Four out of five patients showed good improvementin itching after 3 days in spot B but no improvement in stopping scalingwhich results from cellular hyperproliferation.

Group IV consisting of five patients applied on one selected spot (spotA) control LUBRIDERM cream and on the other selected spot (spot B) creamcontaining 5% retinyl palmitate. No change in spot A was observed after4 weeks of treatment. Five out of five patients showed good improvementin itching after 3 days in spot B but no improvement in stopping scalingwhich results from cellular hyperproliferation.

Group V consisting of five patients applied on one selected spot (spotA) control LUBRIDERM cream and on the other selected spot (spot B) creamcontaining 0.005% calcipotriol. No change in spot A was observed after 4weeks of treatment. Two out of five patients showed partial clearance(average of 35% of spot area) in spot B after 4 weeks of treatment.However, even in the spots showing improvement and partial clearance, acertain amount of scaling is still occurring.

Group VI consisting of five patients applied on one selected spot (spotA) a cream containing 0.0025% calcipotriol and on the other selectedspot (spot B) cream containing 0.0025% calcipotriol and 0.1% retinylpalmitate. One out of five patients showed partial clearance (about 20%of spot area) in spot A after 4 weeks of treatment. However, even inspot A that is showing improvement and partial clearance, a certainamount of scaling is still occurring. Two out of Five patients showedconsiderable amount of clearance (about 40% of spot area) in spot B andwith little scaling and itching.

Group VII consisting of five patients applied on one selected spot (spotA) a cream containing 0.0025% calcipotriol and on the other selectedspot (spot B) cream containing 0.0025% calcipotriol and 0.5% retinylpalmitate. Two out of five patients showed partial clearance (about 25%of spot area) in spot A after 4 weeks of treatment. However, even inspots A that are showing improvement and partial clearance, a certainamount of scaling is still occurring. Three out of Five patients showedconsiderable amount of clearance (about 50% of spot area) in spot B andwith barely noticeable scaling and no itching.

Group VIII consisting of five patients applied on one selected spot(spot A) a cream containing 0.0025% calcipotriol and on the otherselected spot (spot B) cream containing 0.0025% calcipotriol and 1%retinyl palmitate. Two out of five patients showed partial clearance(about 20% of spot area) in spot A after 4 weeks of treatment. However,even in spots A that are showing improvement and partial clearance, acertain amount of scaling is still occurring. Four out of Five patientsshowed considerable amount of clearance (about 50% of spot area) in spotB and with barely noticeable scaling and no itching.

Group IX consisting of five patients applied on one selected spot (spotA) a cream containing 0.0025% calcipotriol and on the other selectedspot (spot B) cream containing 0.0025% calcipotriol and 5% retinylpalmitate. One out of five patients showed partial clearance (about 30%of spot area) in spot A after 4 weeks of treatment. However, even inspot A that is showing improvement and partial clearance, a certainamount of scaling is still occurring. Five out of Five patients showedconsiderable amount of clearance (average about 85% of spot area and onecomplete clearance) in spot B and with no scaling and itching.

Group X consisting of five patients applied on one selected spot (spotA) a cream containing 0.0025% calcipotriol and on the other selectedspot (spot B) cream containing 0.0025% calcipotriol and 10% retinylpalmitate. One out of five patients showed partial clearance (about 15%of spot area) in spot A after 4 weeks of treatment. However, even inspot A that is showing improvement and partial clearance, a certainamount of scaling is still occurring. Five out of Five patients showedconsiderable amount of clearance (average about 90% of spot area and twocomplete clearance) in spot B and with no scaling and itching.

Group XI consisting of five patients applied on one selected spot (spotA) a cream containing 0.0025% calcipotriol and on the other selectedspot (spot B) cream containing 0.0025% calcipotriol and 0.1%4-oxo-retinol. One out of five patients showed partial clearance (about20% of spot area) in spot A after 4 weeks of treatment. However, even inspot A that is showing improvement and partial clearance, a certainamount of scaling is still occurring. Five out of Five patients showedconsiderable amount of clearance (average about 95% of spot area andfour complete clearances) in spot B and with no scaling and itching.

Group XII consisting of five patients applied on one selected spot (spotA) a cream containing 0.1% 4-oxo-retinol and on the other selected spot(spot B) cream containing 0.0025% calcipotriol and 0.1% 4-oxo-retinol.One out of five patients showed partial clearance (about 10% of spotarea) in spot A after 4 weeks of treatment. However, even in spot A thatis showing improvement and partial clearance, a significant amount ofscaling is still occurring. Five out of Five patients showed completeclearances (100%) in spot B and with no scaling and itching.

Group XIII consisting of five patients applied on one selected spot(spot A) a cream containing 0.1% 4-oxo-retinol and on the other selectedspot (spot B) cream containing 0.002% calcitriol and 0.1% 4-oxo-retinol.One out of five patients showed partial clearance (about 15% of spotarea) in spot A after 4 weeks of treatment. However, even in spot A thatis showing improvement and partial clearance, a significant amount ofscaling is still occurring. Five out of Five patients showed completeclearances (100%) in spot B and with no scaling and itching.

Group XIV consisting of five patients applied on one selected spot (spotA) a cream containing 0.1% 4-hydroxy-retinol and on the other selectedspot (spot B) cream containing 0.002% calcitriol and 0.1%4-hydroxy-retinol. One out of five patients showed partial clearance(about 10% of spot area) in spot A after 4 weeks of treatment. However,even in spot A that is showing improvement and partial clearance, asignificant amount of scaling is still occurring. Four out of Fivepatients showed complete clearances with an average clearance of 95% inspot B and with no scaling and itching.

Group XV consists of five patients applied on one selected spot (spot A)a cream containing 0.0025% calcipotriol and on the other selected spot(spot B) cream containing 0.0025% calcipotriol and 0.1% all-transretinoic acid. Two out of five patients showed partial clearance (about15% of spot area) in spot A after 4 weeks of treatment. However, even inspots A that are showing improvement and partial clearance, a certainamount of scaling is still occurring. Similar results were obtained inspot B after 4 weeks of treatment (about 20% partial clearance withpersistence of scaling).

Group XVI consisting of five patients applied on one selected spot (spotA) a cream containing 0.005% calcipotriol and on the other selected spot(spot B) cream containing 5% Cholecalciferol and 5% retinyl palmitate.Two out of five patients showed partial clearance (about 15% of spotarea) in spot A after 4 weeks of treatment. However, even in spots Athat are showing improvement and partial clearance, a certain amount ofscaling is still occurring. Five out of Five patients showedconsiderable amount of clearance (average about 75% of spot area and onecomplete clearance) in spot B and with no scaling and itching. The restof the spots showed no improvement.

This data clearly indicates the synergistic effect of vitamin D analogswith retinyl esters above 0.1%, 4-oxo-retinol, or 4-hydroxy-retinol intreating psoriasis. However, there were no synergistic effects with ofvitamin D analogs with retinoic acid. Similar experiments were carriedout using retinol rather than retinyl palmitate with similar results.

EXAMPLE 3

Three patients diagnosed with eczema were treated with cream containing0.0025% calcipotriol and 5% retinyl palmitate. A significant improvementwas noticed in all three patients within 5 days and two hadnormal-looking skin after two weeks. A similar results was also observedwith patients treated with 0.002% calcitriol and 0.1% 4-oxo-retinol.Treatment with 5% retinyl palmitate, with 0.1% 4-oxo-retinol without avitamin D analog, or with a vitamin D analog and 0.1% all-trans retinoicacid did not result in any significant improvements over the same periodof time.

EXAMPLE 4

A patient diagnosed with atopic dermatitis was treated with creamcontaining 0.05% calcipotriol and 0.1% 4-oxo-retinol. A significantimprovement was noticed in within 1 week and the appearance ofnormal-looking skin after two weeks.

EXAMPLE 5

Various types of human melanoma, breast cancer and prostate cancer cellswill be cultured according to standardized procedures. These cells willbe incubated with in the presence of various concentrations of retinol,retinoic acid, retinyl esters, calcitriol or a combination thereof. Cellgrowth of at least some of these cells will be shown to be significantlyinhibited in the presence of calcitriol and either 4-oxo-retinol or highconcentrations of retinol (about 10⁻⁵ M) as compared with cellsincubated in the absence of the above compounds or in the presence ofeach of the above compounds alone.

EXAMPLE 6

Patients with breast cancer, prostate cancer or leukemia, particularlyacute promyelocytic leukemia treated with a combination of oral doses of4-oxo-retinol or 4-hydroxy-retinol (100 mg/square meter) and oral dosesof calcitriol will have a reduced tumor burden, undergo prolongedremission or are permanently cured.

EXAMPLE 7

Current systemic chemotherapy regimens are unable to prolong survival ofpatients with advanced head and neck cancer. Patients treated with acombination of oral doses of 4-oxo-retinol or 4-hydroxy-retinol (100mg/square meter) and oral doses of calcitriol will survive beyond themedian of 4-6 months, and/or have reduced tumor burden during the periodduring which this treatment is administered.

EXAMPLE 8

Patients with deep (cystic acne) treated with a combination of oraldoses of 4-oxo-retinol or 4-hydroxy-retinol (100 mg/square meter) andoral doses of calcitriol will have greater than 50% mean reduction inlesion counts at the end of 3 to 6 months treatment period and in somecases complete treatment will occur after discontinuation of therapy. Avery prolonged remission and permanent cure for some will be obtained.

EXAMPLE 9

Patients with psoriatic arthritis treated with a combination of oraldoses of 4-oxo-retinol or 4-hydroxy-retinol (50-100 mg/square meter) andoral doses of calcitriol exhibit fewer tender joints and a decreasedduration of morning stiffness.

EXAMPLE 10

Individuals suffering from male pattern baldness treated once or twicedaily with a topical formulation of any of the compositions of thepresent invention, particularly with a topical formulation containing0.002% calcitriol and 0.1% 4-oxo-retinol or 0.1% 4-hydroxy-retinol willregrow a cosmetically significant amount of hair within 6-9 months oftreatment. Similar results would be expected with oral formulations.

EXAMPLE 11

Individuals with gray hair treated once or twice daily with a topicalformulation of any of the compositions of the present invention,particularly with a topical formulation containing 0.002% calcitriol and0.1% 4-oxo-retinal or 0.1% 4-hydroxy-retinol will grow hair having thenatural color within 6-9 months of treatment. Similar results would beexpected with oral formulations.

The invention has been described in terms of preferred embodimentsthereof, but is more broadly applicable as will be understood by thoseskilled in the art. The scope of the invention is therefore limited onlyby the following claims.

What is claimed is:
 1. A composition comprising a vitamin D analog and a retinoid, wherein: (a) the vitamin D analog is capable of binding a vitamin D receptor or being converted in vivo into a compound capable of binding a vitamin D receptor; and (b) the retinoid is selected from the group consisting of retinoid D with an alcohol CH₂OH terminal side chain, an ester of retinoid D having an ester bond, an ether of retinoid D having an ether bond, and retinoid D where the alcohol CH₂OH terminal side chain is replaced with an aldehyde CHO terminal side chain, wherein each of the ester bond and the ether bond is formed with the oxygen at the terminal side chain of Retinoid D and wherein said retinoid D with the alcohol CH₂OH terminal side chain has the structure:

wherein the configuration at the 7-, 9-, 11- and 13-position double bonds is independently Z or E and wherein R₁ is selected from the group consisting of:

wherein the keto group at the 4-position is free or protected; and

wherein X is selected from the group consisting of hydrogen and C₁₋₆-alkyl and Y is selected from the group consisting of hydroxy and C₁₋₆-alkyoxyl and wherein the absolute configuration at the 4-position is independently R or S.
 2. The composition of claim 1, wherein the retinoid is retinoid D with an alcohol CH₂OH terminal side chain.
 3. The composition of claim 1, wherein the retinoid is selected from the group consisting of 4-oxo-retinol, 4-oxo-retinal, and 4-oxo-retinyl ester.
 4. A topical composition according to claim
 1. 5. The composition of claim 1, wherein the vitamin D analog is selected from the group consisting of cholecalciferol, calcifediol, calcitriol, calcipotriol, ergocalciferol, dihydrotachysterol, 1,25-dihydroxyergocalciferol, and 25-hydroxydihydrotachysterol.
 6. The composition of claim 1, wherein the vitamin D analog is calcitriol.
 7. The composition of claim 1, wherein the vitamin D analog is calcipotriol.
 8. The composition of claim 1, wherein the retinoid is 4-oxo-retinol.
 9. The composition of claim 1, wherein the vitamin D analog is calcitriol or calcipotriol and the retinoid is a 4-oxo-retinol.
 10. The composition of claim 1, wherein the vitamin D analog is a vitamin D3 analog.
 11. The composition of claim 1, wherein the retinoid is 4-oxo-retinal.
 12. The composition of claim 1, wherein the retinoid is 4-methoxy-retinol.
 13. A method of treating a subject suffering from a disorder characterized by abnormal cell-proliferation and/or cell-differentiation, comprising administering to the subject in need of such treatment a vitamin D analog and a retinoid, wherein the combination of the vitamin D analog and the retinoid is an effective combination and the retinoid and the vitamin D analog are administered simultaneously or at different times and wherein: (a) the vitamin D analog is capable of binding a vitamin D receptor or being converted in vivo into a compound capable of binding a vitamin D receptor; and (b) the retinoid is selected from the group consisting of retinoid D with an alcohol CH₂OH terminal side chain, an ester of retinoid D having an ester bond, an ether of retinoid D having an ether bond, and retinoid D where the alcohol CH₂OH terminal side chain is replaced with an aldehyde CHO terminal side chain, wherein each of the ester bond and the ether bond is formed with the oxygen at the terminal side chain of Retinoid D and wherein said retinoid D with the alcohol CH₂OH terminal side chain has the structure:

wherein the configuration at the 7-, 9-, 11- and 13-position double bonds is independently Z or E and wherein R₁ is selected from the group consisting of:

wherein the keto group at the 4-position is free or protected; and

wherein X is selected from the group consisting of hydrogen and C₁₋₆-alkyl and Y is selected from the group consisting of hydroxy and C₁₋₆-alkyoxyl and wherein the absolute configuration at the 4-position is independently R or S.
 14. The method of claim 13, wherein the retinoid is retinoid D with an alcohol CH₂OH terminal side chain.
 15. The method of claim 13, wherein the retinoid is selected from the group consisting of 4-oxo-retinol, 4-oxo-retinal, and 4-oxo-retinyl ester.
 16. The method of claim 13, wherein the vitamin D analog is selected from the group consisting of cholecalciferol, calcifediol, calcitriol, calcipotriol, ergocalciferol, dihydrotachysterol, 1,25-dihydroxyergocalciferol, and 25-hydroxydihydrotachysterol.
 17. The method of claim 13, wherein the vitamin D analog is calcitriol.
 18. The method of claim 13, wherein the vitamin D analog is calcipotriol.
 19. The method of claim 13, wherein the vitamin D analog is a vitamin D3 analog.
 20. The method of claim 13, wherein the retinoid is 4-oxo-retinol.
 21. The method of claim 13, wherein the retinoid is 4-hydroxy-retinol.
 22. The method of claim 13, wherein the retinoid is 4-methoxy-retinol.
 23. The method of claim 13, wherein the vitamin D analog is calcitriol or calcipotriol and the retinoid is a 4-oxo-retinol.
 24. The method of claim 13, wherein the retinoid is administered topically.
 25. A method of treating a subject suffering from a disorder selected from the group consisting of psoriasis, acne, atopic dermatitis, eczema, rosacea, actinic keratosis, seborrheic dermatitis, and congenital keratinization disorders, comprising administering to the subject in need of such treatment a vitamin D analog and a retinoid, wherein the combination of the vitamin D analog and the retinoid is an effective combination and the retinoid and the vitamin D analog are administered simultaneously or at different times and wherein: (a) the vitamin D analog is capable of binding a vitamin D receptor or being converted in vivo into a compound capable of binding a vitamin D receptor; and (b) the retinoid is selected from the group consisting of retinoid D with an alcohol CH₂OH terminal side chain, an ester of retinoid D having an ester bond, an ether of retinoid D having an ether bond, and retinoid D where the alcohol CH₂OH terminal side chain is replaced with an aldehyde CHO terminal side chain, wherein each of the ester bond and the ether bond is formed with the oxygen at the terminal side chain of Retinoid D and wherein said retinoid D with the alcohol CH₂OH terminal side chain has the structure:

wherein the configuration at the 7-, 9-, 11- and 13-position double bonds is independently Z or E and wherein R₁ is selected from the group consisting of:

wherein the keto group at the 4-position is free or protected; and

wherein X is selected from the group consisting of hydrogen and C₁₋₆-alkyl and Y is selected from the group consisting of hydroxy and C₁₋₆-alkyoxyl and wherein the absolute configuration at the 4-position is independently R or S.
 26. The method of claim 25, wherein the retinoid is retinoid D with an alcohol CH₂OH terminal side chain.
 27. The method of claim 26, wherein the retinoid is 4-oxo-retinol.
 28. The method of claim 26, wherein the retinoid is 4-hydroxy-retinol.
 29. The method of claim 26, wherein the retinoid is 4-methoxy-retinol.
 30. The method of claim 26, wherein the vitamin D analog is calcitriol or calcipotriol and the retinoid is a 4-oxo-retinol.
 31. The method of claim 25, wherein the retinoid is selected from the group consisting of 4-oxo-retinol, 4-oxo-retinal, and 4-oxo-retinyl ester.
 32. The method of claim 25, wherein the vitamin D analog is selected from the group consisting of cholecalciferol, calcifediol, calcitriol, calcipotriol, ergocalciferol, dihydrotachysterol, 1,25-dihydroxyergocalciferol, and 25-hydroxydihydrotachysterol.
 33. The method of claim 25, wherein the vitamin D analog is calcitriol.
 34. The method of claim 33, wherein the disorder is psoriasis.
 35. The method of claim 33, wherein the disorder is eczema.
 36. The method of claim 25, wherein the vitamin D analog is calcipotriol.
 37. The method of claim 25, wherein the vitamin D analog is a vitamin D3 analog.
 38. The method of claim 25, wherein the disorder is psoriasis.
 39. The method of claim 25, wherein the retinoid is applied topically.
 40. The method of claim 25, wherein the disorder is eczema.
 41. The method of claim 25, wherein the disorder is acne.
 42. The method of claim 25, wherein the disorder is atopic dermatitis.
 43. A method of treating one or more conditions of the skin selected from the group consisting of dry skin, photodamaged skin, age spots, aged skin, inflexibility of stratum corneum, wrinkles, fine lines, actinic blemishes, skin dyschromias, and ichthyosis, comprising applying to the skin having said one or more condition a vitamin D analog and a retinoid, wherein the combination of the vitamin D analog and the retinoid is an effective combination and the retinoid and the vitamin D analog are administered simultaneously or at different times and, wherein: (a) the vitamin D analog is capable of binding a vitamin D receptor or being converted in vivo into a compound capable of binding a vitamin D receptor; and (b) the retinoid is selected from the group consisting of retinoid D with an alcohol CH₂OH terminal side chain, an ester of retinoid D having an ester bond, an ether of retinoid D having an ether bond, and retinoid D where the alcohol CH₂OH terminal side chain is replaced with an aldehyde CHO terminal side chain, wherein each of the ester bond and the ether bond is formed with the oxygen at the terminal side chain of Retinoid D and wherein said retinoid D with the alcohol CH₂OH terminal side chain has the structure:

wherein the configuration at the 7-, 9-, 11- and 13-position double bonds is independently Z or E and wherein R₁ is selected from the group consisting of:

wherein the keto group at the 4-position is free or protected; and

wherein X is selected from the group consisting of hydrogen and C₁₋₆-alkyl and Y is selected from the group consisting of hydroxy and C₁₋₆-alkyoxyl and wherein the absolute configuration at the 4-position is independently R or S.
 44. The method of claim 43, where in the retinoid is retinoid D with an alcohol CH₂OH terminal side chain.
 45. The method of claim 43, wherein the retinoid is selected from the group consisting of 4-oxo-retinol, 4-oxo-retinal, and 4-oxo-retinyl ester.
 46. The method of claim 44, wherein the vitamin D analog is selected from the group consisting of cholecalciferol, calcifediol, calcitriol, calcipotriol, ergocalciferol, dihydrotachysterol, 1,25-dihydroxyergocalciferol, and 25-hydroxydihydrotachysterol.
 47. The method of claim 44, wherein the vitamin D analog is calcitriol.
 48. The method of claim 44, wherein the vitamin D analog is calcipotriol.
 49. The method of claim 44, wherein the retinoid is 4-oxo-retinol.
 50. The method of claim 44, wherein the retinoid is 4-hydroxy-retinol.
 51. The method of claim 44, wherein the retinoid is 4-methoxy-retinol.
 52. The method of claim 44, wherein the vitamin D analog is calcitriol or calcipotriol and the retinoid is a 4-oxo-retinol.
 53. The method of claim 44, wherein the vitamin D analog is a vitamin D3 analog. 